ISOLATION AND PROTEOMIC ANALYSIS OF EXOSOMES SECRETED
BY HUMAN CANCER CELLS IN VITRO
T.A. Shtam,1 S.N. Naryzhny,1 S.B. Landa,1 V.S. Burdackov,2
T.O. Artamonova,2 M.V. Filatov 1,*
1 Petersburg Nuclear Physics Institute, Saint-Petersburg and 2 Saint-Petersburg State Polytechnic University;
* e-mail: fil_53@mail.ru ;
filatov@omrb.pnpi.spb.ru
Exosomes are 20-100 nm membrane vesicles of endocytic origin secreted by most cell types in vitro and in vivo. Since exosomes contain both RNA (mRNA
and microRNA) and proteins, which can be transferred to another cell, and be functional in that new environment, these vesicles may be involved in the communication between
cells. The secretion of exosomes by tumor cells and their implication in the transport and propagation of infectious cargo suggest their participation in pathological situations.
Our purpose here is to describe methods for the production, purification, and proteomic characterization of exosomes derived from human cancer cells in vitro. Based on
exosomes' unique lipidic composition, we have developed the new approach to increase production of exosomes by cells in vitro. Secondly, we have developed quality
control by laser correlation spectroscopy for exosomal assays based on the amount of MHC class I and CD63 molecules on their surface. At last, matrix-assisted laser desorption
ionization time-of-flight (MALDI-TOF) mass spectrometry was used after 2D electrophoresis for the proteomic analysis of exosomes derived from cancer cell lines. This study
describes the protein composition of brain tumor cell-derived exosomes in more detail.
Key words: exosomes, proteomic analysis, laser correlation spectroscopy
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